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Diseases of Aquatic Organisms

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DAO 58:83-87 (2004)  -  doi:10.3354/dao058083

Dot-blot hybridization and RT-PCR detection of extra small virus (XSV) associated with white tail disease of prawn Macrobrachium rosenbergii

Joannes Sri Widada*, Véronique Richard, Zhengli Shi, Dong Qian, Jean-Robert Bonami

Pathogens and Immunity, CNRS, Université Montpellier II, Cc 080, Place Eugène Bataillon, 34095 Montpellier Cedex 5, France

ABSTRACT: The availability of specific and reliable detection methods is essential for monitoring the health status of farmed species, particularly for viral diseases. Extra small virus (XSV), a virus-like particle, is associated with Macrobrachium rosenbergii Noda virus (MrNV) in white tail disease (WTD) of M. rosenbergii. We developed 2 genome-based detection methods for the identification of XSV, namely dot-blot hybridization and a single-step RT-PCR. Detection limits were established and are ca. 2.5 pg and 5 fg of viral RNA for dot-blot hybridization and RT-PCR, respectively. Application of the methods to field samples indicated that some animals positively diagnosed with MrNV did not contain XSV, at least within the detection limit of the methodology. This raises the question of the actual role of XSV and its interactions with MrNV in WTD of M. rosenbergii.

KEY WORDS: Diagnostic tools · Dot-blot hybridization · XSV · MrNV · Macrobrachium rosenbergii · RT-PCR

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